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A Nonhost Peptidase Inhibitor of ~14 kDa from Butea monosperma (Lam.) Taub. Seeds Affects Negatively the Growth and Developmental Physiology of Helicoverpa armigera.

Pandey PK, Singh D, Singh S, Khan MY, Jamal F - Biochem Res Int (2014)

Bottom Line: B. monosperma peptidase inhibitor on 12% denaturing polyacrylamide gel electrophoresis exhibited a single protein band of ~14 kDa with or without reduction.B. monosperma peptidase inhibitor dose for 50% mortality and weight reduction by 50% were 0.5% w/w and 0.10% w/w, respectively.Supplementing B. monosperma peptidase inhibitor in artificial diet at 0.1% w/w, both the efficiencies of conversion of ingested as well as digested food were downregulated, whereas approximate digestibility and metabolic cost were enhanced.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry [DST-FIST & UGC-SAP Supported], Dr. Ram Manohar Lohia Avadh University, Faizabad, Uttar Pradesh 224001, India.

ABSTRACT
Helicoverpa armigera is one of the major devastating pests of crop plants. In this context a serine peptidase inhibitor purified from the seeds of Butea monosperma was evaluated for its effect on developmental physiology of H. armigera larvae. B. monosperma peptidase inhibitor on 12% denaturing polyacrylamide gel electrophoresis exhibited a single protein band of ~14 kDa with or without reduction. In vitro studies towards total gut proteolytic enzymes of H. armigera and bovine trypsin indicated measurable inhibitory activity. B. monosperma peptidase inhibitor dose for 50% mortality and weight reduction by 50% were 0.5% w/w and 0.10% w/w, respectively. The IC50 of B. monosperma peptidase inhibitor against total H. armigera gut proteinases activity was 2.0 µg/mL. The larval feeding assays suggested B. monosperma peptidase inhibitor to be toxic as reflected by its retarded growth and development, consequently affecting fertility and fecundity of pest and prolonging the larval-pupal duration of the insect life cycle of H. armigera. Supplementing B. monosperma peptidase inhibitor in artificial diet at 0.1% w/w, both the efficiencies of conversion of ingested as well as digested food were downregulated, whereas approximate digestibility and metabolic cost were enhanced. The efficacy of Butea monosperma peptidase inhibitor against progressive growth and development of H. armigera suggest its usefulness in insect pest management of food crops.

No MeSH data available.


Related in: MedlinePlus

(a) Purification of BmPI using F2 dialyzed fraction on Sephadex G-75. Each fraction collected through the column is of 4.5 mL at an initial flow rate of 0.3 mL min−1. (b) Trypsin affinity purification of BmPI using fractions (42–58) from Sephadex G-75 and protein profile on 12% SDS-PAGE under reducing conditions. The retained BmPI was eluted with 100 mM HCl. Inset SDS-PAGE of pooled fraction (21 to 36): molecular weight markers are on the left and BmPI from trypsin affinity column chromatography are indicated on the right.
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fig1: (a) Purification of BmPI using F2 dialyzed fraction on Sephadex G-75. Each fraction collected through the column is of 4.5 mL at an initial flow rate of 0.3 mL min−1. (b) Trypsin affinity purification of BmPI using fractions (42–58) from Sephadex G-75 and protein profile on 12% SDS-PAGE under reducing conditions. The retained BmPI was eluted with 100 mM HCl. Inset SDS-PAGE of pooled fraction (21 to 36): molecular weight markers are on the left and BmPI from trypsin affinity column chromatography are indicated on the right.

Mentions: On comparison with other ammonium sulfate precipitated and dialyzed fractions (F1, F3), the fraction F2 exhibited strong inhibitory activity against trypsin and yielded peaks as shown in Figure 1(a) on Sephadex G-75 column. The active fractions (42–58) were pooled, dialyzed, lyophilized, and applied onto a Trypsin-Sepharose CL-4B column. Elution with HCl resulted in a major peak of BmPI with high inhibitory activity against bovine pancreatic trypsin (Figure 1(b)). The purification factor of the purified fraction was 9.1-fold more than that of crude extract with an yield of 7.68% (Table 1). The extracted protein was of ~14 kDa both in the presence and absence of dithiothreitol (Figure 1(b) inset).


A Nonhost Peptidase Inhibitor of ~14 kDa from Butea monosperma (Lam.) Taub. Seeds Affects Negatively the Growth and Developmental Physiology of Helicoverpa armigera.

Pandey PK, Singh D, Singh S, Khan MY, Jamal F - Biochem Res Int (2014)

(a) Purification of BmPI using F2 dialyzed fraction on Sephadex G-75. Each fraction collected through the column is of 4.5 mL at an initial flow rate of 0.3 mL min−1. (b) Trypsin affinity purification of BmPI using fractions (42–58) from Sephadex G-75 and protein profile on 12% SDS-PAGE under reducing conditions. The retained BmPI was eluted with 100 mM HCl. Inset SDS-PAGE of pooled fraction (21 to 36): molecular weight markers are on the left and BmPI from trypsin affinity column chromatography are indicated on the right.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4016866&req=5

fig1: (a) Purification of BmPI using F2 dialyzed fraction on Sephadex G-75. Each fraction collected through the column is of 4.5 mL at an initial flow rate of 0.3 mL min−1. (b) Trypsin affinity purification of BmPI using fractions (42–58) from Sephadex G-75 and protein profile on 12% SDS-PAGE under reducing conditions. The retained BmPI was eluted with 100 mM HCl. Inset SDS-PAGE of pooled fraction (21 to 36): molecular weight markers are on the left and BmPI from trypsin affinity column chromatography are indicated on the right.
Mentions: On comparison with other ammonium sulfate precipitated and dialyzed fractions (F1, F3), the fraction F2 exhibited strong inhibitory activity against trypsin and yielded peaks as shown in Figure 1(a) on Sephadex G-75 column. The active fractions (42–58) were pooled, dialyzed, lyophilized, and applied onto a Trypsin-Sepharose CL-4B column. Elution with HCl resulted in a major peak of BmPI with high inhibitory activity against bovine pancreatic trypsin (Figure 1(b)). The purification factor of the purified fraction was 9.1-fold more than that of crude extract with an yield of 7.68% (Table 1). The extracted protein was of ~14 kDa both in the presence and absence of dithiothreitol (Figure 1(b) inset).

Bottom Line: B. monosperma peptidase inhibitor on 12% denaturing polyacrylamide gel electrophoresis exhibited a single protein band of ~14 kDa with or without reduction.B. monosperma peptidase inhibitor dose for 50% mortality and weight reduction by 50% were 0.5% w/w and 0.10% w/w, respectively.Supplementing B. monosperma peptidase inhibitor in artificial diet at 0.1% w/w, both the efficiencies of conversion of ingested as well as digested food were downregulated, whereas approximate digestibility and metabolic cost were enhanced.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry [DST-FIST & UGC-SAP Supported], Dr. Ram Manohar Lohia Avadh University, Faizabad, Uttar Pradesh 224001, India.

ABSTRACT
Helicoverpa armigera is one of the major devastating pests of crop plants. In this context a serine peptidase inhibitor purified from the seeds of Butea monosperma was evaluated for its effect on developmental physiology of H. armigera larvae. B. monosperma peptidase inhibitor on 12% denaturing polyacrylamide gel electrophoresis exhibited a single protein band of ~14 kDa with or without reduction. In vitro studies towards total gut proteolytic enzymes of H. armigera and bovine trypsin indicated measurable inhibitory activity. B. monosperma peptidase inhibitor dose for 50% mortality and weight reduction by 50% were 0.5% w/w and 0.10% w/w, respectively. The IC50 of B. monosperma peptidase inhibitor against total H. armigera gut proteinases activity was 2.0 µg/mL. The larval feeding assays suggested B. monosperma peptidase inhibitor to be toxic as reflected by its retarded growth and development, consequently affecting fertility and fecundity of pest and prolonging the larval-pupal duration of the insect life cycle of H. armigera. Supplementing B. monosperma peptidase inhibitor in artificial diet at 0.1% w/w, both the efficiencies of conversion of ingested as well as digested food were downregulated, whereas approximate digestibility and metabolic cost were enhanced. The efficacy of Butea monosperma peptidase inhibitor against progressive growth and development of H. armigera suggest its usefulness in insect pest management of food crops.

No MeSH data available.


Related in: MedlinePlus